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The absorbance of calcium standard and calcium were read against blank at 570 nm within 60 minutes.
Samples were calibrated against a calcium standard (10mg/dl, Calcium kit, Sigma and Procedure No. 0150, Stanbio Laboratory), and absorption was measured at 575 ± 5 nm using a spectrophotometer or microplate reader.
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Calcium standards were used for external calibrations in the range of 0.1 10 ppm.
Free calcium concentrations in the reaction medium were determined by following Calcium Green-5N (Molecular Probes, Invitrogen) fluorescence calibrated to calcium standards in reaction medium lacking nitrilotriacetic acid.
By comparison to a laser-cooled calcium optical standard, an upper limit for the fractional frequency instability of 7 x 10 -15 10 -15asured is 1 second of averaging-a value substantially better than that of the world's best measurede atomin clocks.
By comparison to a laser-cooled calcium optical standard, an upper limit for the fractional frequency instability of 7 × 10−15 is measured in 1 second of averaging a value substantially better than that of the world's best microwave atomic clocks.
The Escherichia coli DH5-α [ 2] were transformed with pGFPuv (Clontech, CA, USA) [ 9], by the standard calcium chloride method [ 5].
For miR-206 overexpression or knockout in THP-1 cells, pCDH-CMV-MCS-EF1-copGFP cDNA cloning and expression vector (SBI# CD511B-1, PA, USA) or miRzip lentiviral vector (SBI# MZIPpa/aa-1) was co-transfected with packaging vectors VSVG and PCMV (Clontech, CA, USA) into HEK293T cells using the standard calcium phosphate method.
The Ca1 and Ca6 coordination contacts are consistent with standard calcium bond lengths (2.3 2.5 Å), confirming that both sites bind calcium.
Free EDTA in plasma samples was titrated with a standard calcium chloride solution (50 mM) in the presence of the calcium indicator eriochrome black T (CAS 1787-61-7; 10 mg eriochrome black + 1 g sodium chloride triturated and dissolved in 2.5 mL water).
A standard calcium bath of 1.25 mmol/L was used in CHD and was increased to 1.5 mmol/L when calcium-based binders were reduced or eliminated following conversion to INHD.
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